LAMP detection primers, system and method for detecting health state of exopalaemon carinicauda after WSSV (White Spot Syndrome Virus) infection
A technology of white shrimp, health status, applied in biochemical equipment and methods, recombinant DNA technology, microbial determination/inspection, etc., can solve the problems of fluctuation, increase, decrease, etc. low cost effect
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Embodiment 1
[0041] The screening of the genes related to the health status in the white shrimp and the construction process of the LAMP detection system based on the genes are as follows:
[0042] (1) According to the transcriptome data of white spot syndrome virus (White Spot Syndrome Virus, WSSV) before and after infection of white spot syndrome virus (WSSV), 57 sequences with significant differences in the expression level of transcript levels before and after infection were selected as candidate genes, and primers were designed respectively. The samples before and after WSSV infection were detected by real-time fluorescent quantitative PCR.
[0043](2) According to the results of the preliminary screening, seven genes whose transcription levels changed significantly after infection with pathogens were selected for statistical detection and verification of a single sample. Select 100 cultured healthy white prawns (average body weight, 2.8±0.4g) and raise them temporarily for 3 days. Af...
specific Embodiment 2
[0053] (1) Take approximately 0.2 g gill filaments of shrimps S1 and S2 to be tested and healthy shrimps S3 and S4 respectively, place them in sampling tubes, and grind them quickly with a grinding rod to a slurry state;
[0054] (2) Pick the diaphragm with a toothpick, place it in the sample slurry, and after it is fully wetted, pick it back into the diaphragm tube with a toothpick;
[0055] (3) Absorb absolute ethanol with a straw, drop 2 to 3 drops on the wet sampling diaphragm in the tube, flick the bottom of the tube to stir the diaphragm for 30 seconds;
[0056] (4) Use a new toothpick to transfer the above-mentioned sampling membrane to the rinse tube with the corresponding number, and shake the rinse tube for 3 to 4 minutes;
[0057] (5) Use a new toothpick to transfer the membranes in the above-mentioned rinsing tubes, the positive control FTA membranes in the positive and negative control tubes, and the negative control FTA membranes to the corresponding numbered nuc...
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